Pentobarbital

Xinyue capsules (50 mg PQS/capsule) were provided by JiLin YiSheng Pharmaceutical Co. Ltd. (Jilin, China; SFDA approval certificate number: Z20030073). Enteric-coated aspirin tablets (100 mg) were obtained from Bayer Co. (Leverkusen, Germany; SFDA approval certificate number: J20080078). Clopidogrel tablets were obtained from Sanofi (Paris, France; SFDA approval certificate number: J20080090). Penicillin sodium injection (80 million IU/bottle) was purchased from North China Pharmaceutical Co. Ltd. (Hebei, China; SFDA approval certificate number: X1105313).
Triphenyltetrazolium chloride (TTC) was obtained from Sigma-Aldrich (St. Louis, MO, USA). Adenosine diphosphate (ADP) was obtained from Chrono-Log (Havertown, PA, USA). Fluorescent antibody PE-CD62p and FITC-CD61 were purchased from Bio-Rad Laboratories (Hercules, CA, USA). Thromboxane B₂ (TXB₂), 6-ketone prostaglandin F1α (6-keto-PGF1α), prostaglandin E₂ (PGE₂), tissue plasminogen activator (t-PA), and plasminogen activator inhibitor (PAI) radioimmunoassay kits were obtained from Huaying Engineering of Medicine and Biology Co. Ltd. (Beijing, China). Antibodies against COX-1 and COX-2 were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against VEGF were obtained from Abcam (Cambridge, UK). Pentobarbital was obtained from Sinopharm Chemical Reagents (Beijing, China).

To observe the adaptive responses of mice to long-term exercise, sample collection was performed 36–40 h after the last exercise session in the OME group and the OHE group. The mice in both groups were fasted for 12 h before sample collection to eliminate the effect of exercise-induced stress responses and diet on the various indicators. Each mouse was weighed and subsequently anesthetized by intraperitoneal injection of pentobarbital (50 mg/kg body weight; Sinopharm Chemical Reagent Co., Ltd., Shanghai, China). Blood samples were collected from the orbital venous plexus and centrifuged for 20 min (4°C, 900 g) to separate the serum, which was stored at -80°C before serum testosterone testing. Simultaneously, rapid separation of the testis and epididymis was performed, and the sperm count, motility, and apoptosis rate in the epididymis were measured and assessed [16 (link)]. The separated testis was immersed in liquid nitrogen for rapid freezing and stored at -80°C until further use. The peritesticular, perirenal, and mesenteric fat was separated and weighed on an electronic balance to determine the abdominal fat content of each mouse.

Six SD rats (6–8 weeks old), weighing 200–250 g, provided 24 teeth (upper and lower incisors) for this study and were randomly divided into the control group and IgY@ACP group. After anesthetization with 2% pentobarbital (Sinopharm Chemical, Shanghai, China) intraperitoneal injection, cavities of 0.3 mm depth were prepared in the buccal and cervical regions of rat incisors using a standard bur (Komet, Germany) on a high-speed handpiece with water cooling. Prior to the treatments, all the cavities were treated with 17% EDTA on small cotton pellets, which were changed every 30 s for 15 min, to remove the smear layer. Each tooth was repeated to smear the IgY@ACP slurry daily for 2 min and then rinsed with PBS. The teeth were treated without IgY@ACP as a control group. After daily treatment for four consecutive days, the animals were anesthetized, and the teeth were carefully extracted to avoid damage to their surface. The tooth surface morphology of the rats was also observed by SEM at 5 kV.